The ion microscope is a particularly sensitive analytical tool for the localization of ions such as sodium, potassiuum and calcium which are highly diffusible and of considerable interest in many biological systems. Experiments have been performed that illustrate the necessity of low temperature preparative procedures to eliminate the possibility of ion loss and redistribution of ions not strongly bound to the biological matrix. Several collaborative efforts are being undertaken to apply the currently developed methodology to biological problems in which the transport and compartmentalization of inorganic ions is of importance. Methods of computer digitization and feature analysis can then be applied to the resulting ion images. The ion micrograph negative is digitized and corrected for film emulsion response and quantified in terms of ion intensity. It is proposed to develop biological standard materials well characterized in terms of instrumentally introduced artifacts under different modes of analysis: i.e., ion imaging, ion depth profiling and ion counting. With appropriate standards, individual features in an ion micrograph will be corrected for matrix effects and sputtering artifacts introduced by the analysis process. Digitized biological ion images will then be computer corrected for differing chemical environments within a given image field.